Temesgen BelaynehTemesgen BelaynehMonika Komon-ZelazowskaMonika Komon-ZelazowskaChristian P. KubicekChristian P. Kubicek& Irina S. Druzhinina& Irina S. Druzhinina

IndigenousIndigenous Hypocrea/Trichoderma Hypocrea/Trichoderma species as efficient agents for control species as efficient agents for control of coffee wilt disease in Ethiopiaof coffee wilt disease in Ethiopia

Institute of Chemical Engineering Research Area Applied Institute of Chemical Engineering Research Area Applied Biochemistry and Gene TechnologyBiochemistry and Gene TechnologyGroup of Fungal Evolution and BiodiversityGroup of Fungal Evolution and Biodiversity

Ethiopia as a hotspot for Ethiopia as a hotspot for the global diversitythe global diversity

special country in terms of landraces and biological diversity

a country situated in the Horn of Africa

Many of the plant species are still untouched and still unspoiled

Of the several species of coffee, Ethiopia is the center of origin and diversity of Coffee arabica L.

it accounts for 75% of coffee exports in the world.

Coffea arabicaCoffea arabica L. in Ethiopia L. in Ethiopia

The economy of Ethiopia is based on agriculture and it highly depends on coffee production, therefore coffee is critical to the Ethiopian financial system

Coffee belongs to the botanical family Rubiaceae, which has some 500 genera and over 6,000 species. Most are tropical trees and shrubs which grow in the lower storey of forests

Coffee is one of the most important agricultural commodities in the world, worth up to US$ 14 billion annually

Coffee plant is a large bush Coffee plant is a large bush with dark-green oval leaveswith dark-green oval leaves

•Varieties of Varieties of Coffea arabicaCoffea arabica L.: L.: Typica and BourbonTypica and Bourbon

Coffee wilt Coffee wilt desease desease

(tracheomycosis)(tracheomycosis)

Currently, the Ethiopian national average of coffee yield is about 472 Kg ha-1. That is much below the research results of 1500-2000 Kg ha-1.

Such a substantial loss is to large extend determined by the wide spread of coffee wilt diseases which are usually caused by Fusarium species

Coffee wilt Coffee wilt desease desease

(tracheomycosis)(tracheomycosis)Before its occurrence in Ethiopia these fungi were earlier reported to be well known pathogen of other coffee species in the West and Central Africa attacking mostly Coffee excelsa plantations especially in Cameroon and later C. canephra (Robusta coffee) in Ivory Coast and Zaire. Its occurrence in Ethiopia was identified in the early 1970’s when Kranz and Mogk (1973) isolated pure culture from dying coffee trees.

Established in 1974 by the former USSRwith the science and technology development agreement

Ethiopian Plant Protection Ethiopian Plant Protection Research Center Research Center

Sections:Sections:

MycologyBacteriologyVirologyNematology Entomology Departmentsand Biocontrol is a section within biotechnology

Ethiopian Plant Protection Ethiopian Plant Protection Research Center Research Center

MSc Temesgen Balayneh MSc Temesgen Balayneh

Coffee projectCoffee project

Ethiopian coffee ceremonyEthiopian coffee ceremony

Coffee wilt disease and type of forestry

making of artificial coffee „plantations“ by desruption of original forest, cultivation of fast growing shadow trees and subsequent introduction of coffee plants

introduction of individual plants of C. arabica in the natural forest ecosystem

Resistance to coffee wilt Resistance to coffee wilt disease (hypothesis)disease (hypothesis)

due to presence of native antagonists in rhizosphere of C. arabica

due to symbiotic fungi (Glomus spp. and ?? Trichoderma ???)

due to multidimentional interactions between coffee plant, symbionts and rhizosphere microbionts

Goals of the research Goals of the research

to study the native biodiversity of Trichoderma in Ethiopia

to identify the causative agent of coffee wilt disease (Fusarium spp.)

to test the potential of different Trichoderma isolates from rhizosphere of Coffea arabica L. to control Fusarium

Road side survey Road side survey

Jimma

Harerga

SNNP

Wellega

•With 10 to 15 km stop of carWith 10 to 15 km stop of car

•Collected in plastic bagsCollected in plastic bags

•Stored at -4°C until processingStored at -4°C until processing

•Pilot project for diversity studyPilot project for diversity study

Fusarium diversity

strains of Fusarium causing coffee wilt disease were isolated directly from infected roots

based of tef1 sequence analysis using FUSARIUM-SEQ (Geiser et al., 2004) we identified them as Gibberella fujikuroi/ Fusarium oxysporum species complex:

TrichodermaTrichoderma biodiversity biodiversity

• Generally, we collected 60 soil samples

- Different coffee growing areas

- Diff. Cultivation:- Forest, Semi-forest and “Plantation”

- Alt. Range:- 1300 - 2400 m.a.s.l.

• With the visual and microscopic identification

- Found 45 samples of Trichoderma spp.

TrichodermaTrichoderma biodiversity biodiversity

All Trichoderma strains have been identified based on ITS1 and 2 and tef1 sequence analyses using TrichOKEY and TrichoBLAST tools, respectively (www.isth.info)

TrichodermaTrichoderma biodiversity biodiversity

List of species: Amount Strain numbers in CPK collection:

T. harzianum 5 1807, 1812, 1818, 1825, 1840

T. brevicompactum 1 1828

T. oblongisporum 1 1809

T. spirale 2 1822, 1839

T. hamatum 5 1810, 1811, 1814, 1826, 1827, 1835

T. koningiopsis 5 1813, 1816, 1821, 1823, 1831

T. asperellum 4 1819, 1820, 1829, 1838

T. atroviride 1 1832

T. longibrachatum 2 1817, 1889

T. orientalis 2 1815, 1837

New species 91808, 1833, 1834, 1836, 1841, 1888, 1890, 1892, 1893

TrichodermaTrichoderma diversity in diversity in C. arabicaC. arabica rhizosphere rhizosphere

TrichodermaTrichoderma biodiversity in Ethiopia biodiversity in Ethiopia (coffee growing forest)(coffee growing forest)

New species 1, Sect. Longibrachiatum, related to T. longibrachiatum - H. orientalis species complex

New species 2, Sect. Pachybasium, related to T. tomentosum - T. cerinum from T. harzianum species complex

New species 3Sect. Trichoderma, related to T. asperellum

New species 4Sect. Pachybasium, related to T. spirale

Jimma

Welega Jimma

Harerga SNNP

Jimma

JimmaNew species 5Sect. Pachybasium, related to T. helicum

TrichodermaTrichoderma biodiversity in Ethiopia biodiversity in Ethiopia (coffee growing forest)(coffee growing forest)

TrichodermaTrichoderma biodiversity in Ivory Coast biodiversity in Ivory Coast (coccoa plantations)(coccoa plantations)

in vitroin vitro tests of antagonistic tests of antagonistic potentialpotential

• Fusarium – Trichoderma confrontations on plates

• Ability of Trichoderma to overgrow Fusarium cultures

experimental strategies

• P1 strain of Trichoderma atroviride as the reference

in vitroin vitro tests of antagonistic tests of antagonistic potentialpotential

List of species: Amount Numbers from CPK collection Jimma Harerga SNNP Wellega

T. harzianum 5 1807, 1812, 1818, 1825, 1840                                

T. brevicompactum 1 1828                                

T. oblongisporum 1 1809                                

T. spirale 2 1822, 1839                                

T. hamatum 6 1810, 1811, 1814, 1826, 1827, 1835                                

T. koningiopsis 5 1813, 1816, 1821, 1823, 1831                                

T. asperellum 5 1819, 1820, 1829, 1838                                

T. atroviride 1 1832                                

T. longibrachatum 1 1817                                

T. orientalis 2 1815, 1837                                

New species 1 3 1808, 1833, 1892                                

New species 2 2 1836, 1893                                

New species 3 2 1841, 1890                                

New species 4 1 1888                                

New species 5 1 1834                                

strongstrong moderatemoderate weakweak no responseno response

T. oblongisporum T. oblongisporum (Jimma)(Jimma)

T. koningiopsisT. koningiopsisSamuels et al., ms submitted to Stud. Myc.Samuels et al., ms submitted to Stud. Myc.

T. atroviride T. atroviride (Jimma)(Jimma)

T. T. sp. 1sp. 1 (Jimma and SNNP)(Jimma and SNNP)

T. T. sp. 2sp. 2 (Welega and SNNP)(Welega and SNNP)

ConclusionsConclusions

The first road side survey study has revealed 10 known and 5 potentially new species of Trichoderma inhibiting the rhizosphere of Coffea arabica. The biodiversity of Trichoderma is very high.

In all four areas coffee wilt disease is caused by strains of Fusarium oxysporum species complex. The biodiversity of Fusarium is low.

25 out of 45 (55%) Trichoderma strains have strong potential to prevent the growth of Fusarium oxysporum.

Future perspectivesFuture perspectives

AknowledgementsAknowledgements

The research was supported by Ethiopia:The research was supported by Ethiopia: PPRC scholarship to Temesgen BalaynehPPRC scholarship to Temesgen Balayneh

and partly by FWF projectand partly by FWF project

Thank you for your attention!Thank you for your attention!